Molecular identification of Taenia specimens after long-term preservation in formalin

• Published in: Parasitology international 2011-06, Vol.60 (2), p.203-205
• Main Authors: Jeon, Hyeong-Kyu, Kim, Kyu-Heon, Eom, Keeseon S
• Format: Article
• Language: English
• Subjects: Animals; DNA; DNA, Helminth - chemistry; DNA, Helminth - genetics; DNA, Helminth - isolation & purification; Endopeptidase K - metabolism; Fixatives - chemistry; Formaldehyde - chemistry; formalin; Formalin fixed museum collection; Gastroenterology and Hepatology; genes; Genes, Helminth; Genome, Helminth; Indonesia; Infectious Disease; Japan; Molecular identification; natural history; nitrogen; nucleotide sequences; parasitology; peptidase K; polymerase chain reaction; Polymerase Chain Reaction - methods; Preservation, Biological; Republic of Korea; Sequence Analysis, DNA - methods; storage time; Taenia; Taenia - classification; Taenia - genetics; tapeworms; Tissue Fixation
• ISSN: 1383-5769; 1873-0329
• DOI: 10.1016/j.parint.2010.12.001

Abstract The majority of Taenia tapeworm specimens in the museum collections are usually kept in a formalin fixative for permanent preservation mainly for use in morphological examinations. This study aims to improve Taenia tapeworm identification even of one preserved in formalin for a maximum of 81 years. Taenia tapeworms were collected by the parasite collection unit of the Swiss Natural History Museum and from units in Indonesia, Japan and Korea. A small amount of formalin-fixed tissue (100 mg) was crushed in liquid nitrogen and then soaked in a Tris–EDTA buffer for 3–5 h. The sample was then digested in SDS and proteinase K (20 mg/ml) for 3–5 h at 56 °C. After the addition of proteinase K (20 mg/ml), SDS and hexadecyl-trimethyl-ammonium bromide (CTAB), incubation was continued for another 3 h at 65 °C. A maximum yield of genomic DNA was obtained from this additional step and the quality of genomic DNA obtained with this extraction method seemed to be independent of the duration of storage time in the formalin fixative. The molecular identification of Taenia tapeworms was performed by using PCR and DNA sequences corresponding to position 80–428 of cox1 gene. T. asiatica was detected in the isolates of Indonesia, Japan and Korea. Improvements in the genomic DNA extraction method from formalin fixed museum collections will help in the molecular identification of parasites.